*Target (protein/gene name): protein kinase, putative
*NCBI Gene # or RefSeq#: 3530995
*Protein ID (NP or XP #) or Wolbachia#:Tc00.1047053398235.10
*Organism (including strain):Trypanosoma cruzi
Etiologic Risk Group (see link below): Risk Group 2 (RG2) - Parasitic Agents
*Background/Disease Information (sort of like the Intro to your Mini Research Write up): T cruzi is a species of parasitic euglenoid trypanosomes. It causes trypanosomiasis in humans and animals. They trasmit through feces and bites. Also known as Chagas disease, one form is found in blood and another in tissues. The life cycle begins in a reservoir of mammals, through the triatomine bug. They can swim into host cells using flagella. They then reproduce through binary fission until the cell bursts.
Essentiality: This protein adds phosphate groups causing a functional change, up to 30% of human proteins are modified.
Complex of proteins?: works by itself.
Druggable Target: Druggability index (range: 0 to 1): 0.1
Structure Available (PDB or Homology model)
-- PDB # or closest PDB entry if using homology model: 4GVY
Current Inhibitors:
2-oxoglutarate
5,5'-dithiobis(2-nitrobenzoic acid)
agmatine
Aminoguanidine
canavanine
dithiothreitol
Guanidine hydrochloride
Expression Information (has it been expressed in bacterial cells): It has been expressed in Escherichia coli BL21 (DE3) cells
Purification Method: expressed as as a fusion with maltose-binding protein
Image of protein (PyMol with features delineated and shown separately):
*
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):
*length of your protein in Amino Acids 357
Molecular Weight of your protein in kiloDaltons using the Expasy ProtParam website 40.1937 kDa
Molar Extinction coefficient of your protein at 280 nm wavelength: 33140
TMpred graph Image (http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it.
Do Not Need this info for Spring (but still copy these lines to your Target page for now)
Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers):
(link to DNA Works output text file - that should be saved in your Google Docs folder after you did the primer design protocol)
-- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to.
Primer design results for 'tail' primers (this is just 2 sequences):
*NCBI Gene # or RefSeq#: 3530995
*Protein ID (NP or XP #) or Wolbachia#:Tc00.1047053398235.10
*Organism (including strain):Trypanosoma cruzi
Etiologic Risk Group (see link below): Risk Group 2 (RG2) - Parasitic Agents
*Background/Disease Information (sort of like the Intro to your Mini Research Write up): T cruzi is a species of parasitic euglenoid trypanosomes. It causes trypanosomiasis in humans and animals. They trasmit through feces and bites. Also known as Chagas disease, one form is found in blood and another in tissues. The life cycle begins in a reservoir of mammals, through the triatomine bug. They can swim into host cells using flagella. They then reproduce through binary fission until the cell bursts.
Essentiality: This protein adds phosphate groups causing a functional change, up to 30% of human proteins are modified.
Complex of proteins?: works by itself.
Druggable Target: Druggability index (range: 0 to 1): 0.1
*EC#: 2.7.3.3
Link to BRENDA EC# page:
http://www.brenda-enzymes.info/literature/lit.php4?e=2.7.3.3&r=642469
-- Show screenshot of BRENDA enzyme mechanism schematic
Enzyme Assay information (spectrophotometric, coupled assay ?, reagents):
-- link to Sigma (or other company) page for assay or assay reagents (substrates)
http://www.sigmaaldrich.com/life-science/metabolomics/enzyme-explorer/learning-center/assay-library/ec-number-ii.html
-- link (or citation) to paper that contains assay information
-- List cost and quantity of substrate reagents and supplier
http://www.brenda-enzymes.info/literature/lit.php4?e=2.7.3.3&r=642482
Structure Available (PDB or Homology model)
-- PDB # or closest PDB entry if using homology model: 4GVY
Current Inhibitors:
2-oxoglutarate
5,5'-dithiobis(2-nitrobenzoic acid)
agmatine
Aminoguanidine
canavanine
dithiothreitol
Guanidine hydrochloride
Expression Information (has it been expressed in bacterial cells): It has been expressed in Escherichia coli BL21 (DE3) cells
Purification Method: expressed as as a fusion with maltose-binding protein
Image of protein (PyMol with features delineated and shown separately):
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):
*length of your protein in Amino Acids 357
Molecular Weight of your protein in kiloDaltons using the Expasy ProtParam website 40.1937 kDa
Molar Extinction coefficient of your protein at 280 nm wavelength: 33140
TMpred graph Image (http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it.
*CDS Gene Sequence (paste as text only):
*GC% Content for gene:
GC content 48.4%
Do Not Need this info for Spring (but still copy these lines to your Target page for now)
Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers):
(link to DNA Works output text file - that should be saved in your Google Docs folder after you did the primer design protocol)
-- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to.
Primer design results for 'tail' primers (this is just 2 sequences):